Anti-Müllerian hormone (AMH) is a homodimeric glycoprotin secreted by granulosa cells in postnatal females. AMH plays an important role in follicle recruitment. The aim of the study was to analyse AMH concentrations under the guidance of circulating estradiol and progesterone levels in mares. The study was conducted on 25 non-lactating mares with regular estrous cycles. Blood samples were collected during estrus and diestrus to analyse estradiol, progesterone and AMH concentrations. Estradiol and progesterone concentrations were significantly different (P < 0.001) between estrus and diestrus. AMH concentrations in estrus were significantly higher than those in diestrus (P < 0.05). According to the authors’ knowledge, this is the first report to suggest that the sexual cycle stage affects AMH concentrations in mares. Further investigations are needed to reveal factors affecting the AMH production and secretion pattern throughout the sexual cycle.